Biomedical subjects

P C Brunjes

Publications and source records attributed to P C Brunjes.

At least 19 recordsLinked to original sources

PubMed1995

Organization of the olfactory system in the adult zebrafish: histological, immunohistochemical, and quantitative analysis.

The zebrafish, Danio rerio, is becoming an important model system for developmental studies. We have used a variety of histological techniques to characterize the adult structure of the olfactory system in this teleost to form a base for future developmental work. The olfactory epithelium in this fish contains ciliated and microvillar sensory neurons, microvillar supporting cells, secretory goblet cells, and basal cells, and the adjacent nonsensory epithelium contains ciliated supporting cells. The olfactory bulb is a diffusely organized structure with four laminae: olfactory nerve, glomerular, mixed mitral cell/plexiform, and granule cell layers. These structures and the synapses observed in the olfactory bulb are typical of what is found in other vertebrates. We also examined the distribution of several neurotransmitter markers (tyrosine hydroxylase, neuropeptide Y, dopamine-beta-hydroxylase, and serotonin) in the olfactory bulb. Antibodies to neuropeptide Y, dopamine-beta-hydroxylase, and serotonin labeled fibers in the olfactory bulb and cell bodies in caudal regions of the brain in distributions comparable to other species. Tyrosine hydroxylase immunoreactivity was observed in a set of intrinsic bulb neurons with extensive processes in the glomerular layer. In addition, the structural proteins glial fibrillary acidic protein and vimentin have distributions similar to those in the olfactory bulbs of other animals. Thus, the adult olfactory structures are analogous to the structures in other vertebrate animals in morphology and chemical neuroanatomy. This similarity, along with its numerous advantages for developmental studies, makes the zebrafish a good model for studies of olfaction and forebrain maturation.

Animals↗

PubMed1995

Fos protein immunoreactivity in the developing olfactory bulbs of normal and naris-occluded rats.

Immediate early genes such as c-fos may be a route through which extracellular events affect genomic expression. Expression of immediate early genes is important in the transcriptional regulation necessary for the normal development of the nervous system. Developmental patterns of Fos protein (the product of c-fos immediate early gene expression) were studied in the main olfactory bulb of the rat using immunocytochemistry. Embryonic Day 21 (E21, the last prenatal day), as well as Postnatal Day 0 (P0), P1, P5, P10, P15, P20 and P30 subjects were examined. Although staining was absent in the E21 bulb, there was a rapid onset of Fos synthesis within hours after birth. Distribution of Fos-immunoreactive (Fos-ir) nuclei corresponded to the sequence of bulb maturation: numerous mitral/tufted and granule cells were labeled on P0, followed by the appearance of Fos-ir in the nuclei of periglomerular cells and an increase in the number of stained granule cells with development. Surgical closure of an external naris on P1 resulted in a 70% reduction in the number of Fos-ir granule cell nuclei as early as 2 h after the manipulation. During the next 30 days, levels of Fos staining further diminished in experimental bulbs when compared to their contralateral controls. Nevertheless, electrical stimulation of the contralateral bulb in P20 pups resulted in a robust increase of Fos labeling in most main and accessory olfactory bulb mitral cells and in many granule and periglomerular neurons, suggesting that the experimental bulbs remain competent to express Fos protein.

Aging↗

PubMed1995

Intracellular injections of lucifer yellow into lightly fixed mitral cells reveal neuronal dye-coupling in the developing rat olfactory bulb.

We report that intracellular injections of Lucifer Yellow into lightly fixed mitral cells revealed dye-coupling between mitral cells and between mitral and granule cells in the form of discrete, radially oriented cell clusters. Dye-coupling was observed in animals as early as postnatal day 10 (P10) and at least until P30. In P10 rats, a mean of 2.5 dye-coupled mitral cells and 27 granule cells were observed per column. Mean column depth and width were 169 microns and 86 microns respectively. Most of the dye-filled granule cells were found within 150 microns of the mitral cell layer. No significant changes were found at P20. By P30, the mean number of granule cells per column increased to 42 and the addition of granule cells occurred in areas proximal to the mitral cell layer. Immunocytochemical results indicate that the developing bulb contains a large concentration of the gap junction protein Connexin 43 (Cx43). Cx43-like immunoreactivity was found at all ages examined, with the most intense staining in the nerve and glomerular layers. Less intense Cx43-like immunoreactivity was found in both the mitral and granule cell layers, with Cx43-like immunoreactive puncta observed between and around cell body profiles. Freeze-fracture analysis revealed the presence of gap junction-like plaques on mitral cells, further suggesting that the dye-coupling occurred across interneuronal gap junctions. Neuronal coupling during development could provide an inter-cellular pathway for the passage of relevant developmental signals which could influence the formation and/or strengthening of synaptic contacts. The coupling could also be involved in the synchronization of neuronal activity, which may be important for olfactory coding.

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PubMed1995

Oligodendrocyte/myelin-immunoreactivity in the developing olfactory system.

Immunocytochemistry was used to characterize oligodendrocyte maturation in the developing mammalian olfactory system. Postnatal day 10-16, 20, 30 and adult rats were examined, as well as postnatal day 20, 30, 40 and adult Monodelphis domestica (the grey, short-tailed opossum). In rats, oligodendrocyte/myelin-immunoreactivity first appears in the accessory olfactory bulb by day 11, with labeling rapidly increasing throughout the entire bulb over the next five days. An adult pattern of immunoreactivity, characterized by dense labeling in the granule cell layer, sparse immunoreactivity in the external plexiform layer, and staining along the periphery of glomeruli, is attained by day 30. Staining is apparent in both the lateral olfactory tract and anterior commissure by day 11, and becomes heavy by day 20. While patterns of oligodendrocyte/myelin-immunoreactivity in the adult Monodelphis and rat bulb are similar, staining first appears much later in the opossum (around day 30), and maturation occurs more slowly. For example, rostral-caudal gradients in the development of staining in the anterior commissure were noted which were not seen in the rat. These differences emerge because Monodelphis' slower growth allows more resolution into developmental sequences. Finally, in rats, unilateral naris closure on the day after birth, which significantly alters normal patterns of bulb development, has no effect on the pattern and level of immunoreactivity even after long (30 day) survival periods. In both normal and naris occluded rats, oligodendrocyte/myelin-immunoreactivity is found in caudal aspects of the rat bulb on day 11 and subsequently progresses throughout the entire bulb over the next five days. Patterns in the Monodelphis bulb mirror those observed in the rat, however, staining appears later and progresses more slowly, suggesting Monodelphis is a useful animal for examining early myelin formation.

Age Factors↗

PubMed1994

The development of serotonergic projections to the olfactory bulb of Monodelphis domestica (the grey, short-tailed opossum).

The offspring of Monodelphis domestica (grey, short-tailed opossum) are born in a very immature state after a short (14-day) gestation period. As a result, they provide a useful mammalian model for examining quite early stages of brain maturation. The present study demonstrates that the onset and development of serotonergic (5-HT) projections to the Monodelphis olfactory bulb can be traced entirely after birth. 5-HT was visualized in tissue sections from postnatal day 5 (P5), P10, P20, P30, and adult opossums using avidin-biotin peroxidase immunocytochemistry. 5-HT afferent fibers are rare in the bulb until P10. Maturation occurs slowly, but by P30, 5-HT fibers assume the adult form characterized by preferential glomerular innervation. Due to the postnatal development, slow maturation, and specificity of innervation patterns, the Monodelphis bulb provides a model with which to study the regulation of serotonergic development.

Aging↗

PubMed1994

Changes in cell proliferation in the developing olfactory epithelium following neonatal unilateral naris occlusion.

Surgical closure of an external naris reduces airflow through one side of the nasal cavity. Previous studies using [3H]thymidine autoradiography have demonstrated that rats subjected to the procedure on Postnatal Day 1 (P1) exhibit reduced cellular proliferation in the olfactory mucosa by P30. We investigated cell production at earlier ages to determine (a) when the effect first emerges, and (b) whether septal, dorsal, and lateral regions of the nasal cavity responded similarly. Both short (2-h) and long (30-day) survival groups were tested. Results from the first group indicate that changes in proliferation emerge between P10 and P20, and that the septum is more affected than other regions of the nasal cavity. No differences were observed in the long survival groups, perhaps due to changes in patterns of cell production or death.

Airway Obstruction↗

PubMed1994

Unilateral naris closure and olfactory system development.

In most animals there is bilateral access of odorants to the olfactory sensory epithelium. Air enters the nose through two external nares and passes back through the nasal cavity, which is divided down the midline by a cartilaginous nasal septum. The olfactory mucosa, a sheet of ciliated bipolar receptor cells, is found in the caudal two thirds of the nasal cavity. Axons from the sensory cells project to an ipsilateral extension of the telencephalon known as the olfactory bulb. If a single external naris of a rat pup is surgically closed (usually via brief cauterization) on the day after the day of birth (P1) and the subject is examined on P30, the size of the ipsilateral olfactory bulb is reduced by approximately 25%. The large reduction in size, coupled with the clear lamination and other features of the olfactory system, indicates that the manipulation is an ideal preparation for examining the regulation of early growth. We know that both olfactory bulbs are of equal size at the time of occlusion, but that 30 days later there is a large discrepancy. What series of events produces the changes? The present paper outlines what is known about the anatomical, biochemical and physiological changes introduced by naris occlusion in order to lay a framework for further work.

Animals↗

PubMed1993

Spatial organization and plasticity of the primary and secondary olfactory projections in goldfish.

Crystals of the lipophilic tracer DiI were applied to discrete regions of the olfactory epithelium of goldfish to trace the primary sensory projection to the olfactory bulb. Receptors from the anterior half of the sensory sheet project primarily to glomeruli in the medial half of the bulb and receptors in the posterior half terminate mainly within the lateral half of the bulb. This pattern disappeared following ablation of selected, discrete epithelial regions. In order to investigate reorganization of secondary olfactory projections, unoperated control and unilaterally bulbectomized animals received injections of [3H]proline into the right olfactory bulb. Densities of silver grains per unit area were determined within six different forebrain nuclei in both the right and left hemispheres of each animal. Of the six areas examined, three demonstrated a significantly greater density of afferent innervation from the ipsilateral versus contralateral bulb; a difference which disappeared in two of these three regions after bulbectomy. Thus, for at least two forebrain nuclei, bulb removal caused a change in the afferent input from the spared olfactory bulb to those regions. We conclude that both primary and secondary olfactory projections in goldfish are capable of some degree of reorganization following insult.

Animals↗

PubMed1993

Microglia and the developing olfactory bulb.

The developmental appearance of microglia in the rat olfactory bulb was investigated through the use of selective staining with the B4-isolectin from Griffonia simplicifolia. No changes in the density or distribution of either the spherical, macrophage "ameboid" form or the highly arborized "ramified" variety of microglia were observed in the superficial layers of the bulb between postnatal days 10 and 30. The subependymal zone exhibited the only substantial population of ameboid cells and the only developmental increases in ramified cell density during this time-period. External single naris closure, which enhances cell death in the ipsilateral bulb, did not affect microglia density, presumably due to the unusually high numbers of microglia normally present in the bulb. The olfactory bulb has a dense and relatively uniform population of microglial cells from very early stages of postnatal life, perhaps because of the constant turnover of cells and processes.

Aging↗

PubMed1992

Olfactory bulb organization and development in Monodelphis domestica (grey short-tailed opossum).

The olfactory bulbs of adult and developing Monodelphis domestica were examined with a number of techniques. Golgi, Nissl, and Timm stains as well as acetylcholinesterase histochemistry revealed a high degree of order within the adult bulb. All major cell classes characteristic of most mammalian species were observed. Tufted cells appeared to be restricted to the superficial portion of the external plexiform layer. Developing Monodelphis pups were examined with Nissl-stained semithin sections and with immunocytochemistry for tyrosine hydroxylase, microtubule-associated protein 2, vimentin, and glial fibrillary acidic protein. Newborn pups are extremely immature, with few postmitotic cells present in the forebrain. Considerable maturation occurs over the first four postnatal weeks, and by postnatal day 30, the bulb assumes an adult-like organization. The extreme immaturity of the bulb at birth, coupled with its strict organization, suggest that Monodelphis is a particularly appropriate species for experimental examinations of olfactory system development.

Acetylcholinesterase↗

PubMed1992

Unilateral naris closure and vascular development in the rat olfactory bulb.

The blood supply to the brain has been linked closely to nervous system function and metabolism, thereby possibly playing a direct role in brain maturation. Previously, we demonstrated that closure of an external naris early in life results in large changes within the olfactory bulb, including reductions in laminar volume and cell number and a rapid decline in metabolism and protein synthesis. To understand the role of the blood supply in the dramatic changes following naris closure, the present study examines the development of olfactory bulb vasculature in unilaterally odor-deprived and control rats. On post-partum day 1 (P1; the day after birth), littermate rat pups underwent either unilateral naris occlusion or sham surgery. On P5, P10, P15, P20, P30 and P60, animals were perfused with an india ink-gelatin mixture to assess blood vessel amount and complexity. Densitometric analyses were performed to obtain values of blood vessel area ratios (vessel area/tissue area), branch point number and branch point density. Considerable vessel development in all bulbs occurred over the first two to three weeks post-partum. By P20, large reductions in vessel area ratios were observed in all constituent laminae of deprived bulbs. While similar reductions in number of vessel branch points/tissue area were seen, few changes were noted in the number of branch points/vessel area. The effects were primarily confined to early developmental periods: bulb vasculature in animals deprived at older ages (P40) appeared normal. The results indicate that the vasculature responds to alterations in sensory stimulation early in life, therefore potentially playing an important regulative role in neural development.

Aging↗

PubMed1991

Unilateral olfactory deprivation: effects on succinate dehydrogenase histochemistry and [3H]leucine incorporation in the olfactory mucosa.

Surgically closing one external naris reduces airflow through one half of the nasal cavity, decreasing the access of odors to the receptor sheet. In rats, unilateral naris occlusion performed near birth results in large reductions in the size of the olfactory bulb, the primary central relay, when examined 30 days later. Previous research has demonstrated that there is a rapid reduction in [3H]2-deoxyglucose (2-DG) and [3H]leucine uptake in the bulb within hours after naris closure. The present study examined whether similar rapid changes could be observed in the sensory periphery. Pups occluded on P1 and examined on P3 with succinate dehydrogenase histochemistry exhibited reduced staining on the closed side of the nasal cavity, suggesting occlusion results in reductions in mucosal metabolism. Larger differences in staining were observed in pups examined at P6. [3H]Leucine incorporation was quite similar on both sides of the nasal septum as late as 30 days post occlusion, suggesting less dramatic changes in protein synthesis. The results suggest that naris closure does indeed have rapid effects on mucosal function, but indicate that the changes are different than those observed in the bulb.

Animals↗

PubMed1990

Development of the anterior olfactory nucleus in normal and unilaterally odor deprived rats.

The development of a second order structure in the olfactory pathway, the anterior olfactory nucleus, was examined in both normal rat pups and in subjects which underwent unilateral naris closure on postnatal day 1 (P1). Naris occlusion in neonatal rats produces a constellation of changes within the first relay in the pathway, the olfactory bulb, including a 25% reduction in total volume. Such large changes suggest that higher order structures might also be affected. Anterior olfactory nucleus development was quantified in several ways. Laminar volumes were computed by using serial section planimetry. In control animals differential development was observed, with regions extending most rostrally (e.g., pars externa and pars lateralis) exhibiting the least growth. The anterior olfactory nucleus on the "deprived" side of subjects with a single naris occluded was identical in size to that observed in controls, development within the pars lateralis was examined in control animals at P10, P20, P30, and adults. Developmental increases in numbers of both branches per cell and spines were noted, but mean branch length remained relatively constant. Finally, the effects of naris occlusion on histological patterns of succinate dehydrogenase (SDH) staining and 2-deoxyglucose uptake within pars lateralis were examined at P20 to test for more subtle effects of naris occlusion. SDH staining was quite similar in deprived and control rats at P20. However, 3H-2-DG uptake was decreased in rostral areas of the anterior olfactory nucleus ipsilateral to the deprived olfactory bulb, suggesting that naris closure does affect the structure.

Animals↗

PubMed1990

Olfactory bulb and sensory epithelium in goldfish: morphological alterations accompanying growth.

Considerable growth of both the body and the brain occurs throughout the lifespan in goldfish. In the present study growth-related changes in the goldfish olfactory system were examined using quantitative morphometric techniques as well as [3H]thymidine autoradiography. While the two olfactory structures examined, the bulb and the rosette, grow in proportion to the rest of the animal, each structure shows evidence of differential maturation. Examination of olfactory bulb laminar volumes suggest that bulbs expand primarily by generating neuropil rather than by adding new cells. This interpretation was supported by the findings that olfactory bulb granule cells decrease in density and increase in soma size as animals grow. Additionally, virtually no [3H]thymidine-labeled cells were found in the granule cell zone. Olfactory rosettes grow by adding new lamellae to the structure's medial aspect as well as by increasing the length of existing lamellae. These findings were supported by examining [3H]thymidine-labeled cells, which were differentially distributed across the rosette, with significantly more labeled cells found medially vs laterally. Results are discussed in terms of their implications for the system's functioning during growth.

Animals↗

PubMed1990

Rapid changes in 2-deoxyglucose uptake and amino acid incorporation following unilateral odor deprivation: a laminar analysis.

Unilateral naris occlusion in neonatal rats results in large alterations in the olfactory bulb, including substantial changes in laminar volume and enhanced cell death. These gross changes are undoubtedly the result of a cascade of more basic cellular regulatory events. The present study assesses the possibility of rapid post-deprivation changes in two such processes: glucose metabolism and protein synthesis. On the day after the day of birth rat pups underwent unilateral naris occlusion or sham surgery. In one study, either 1, 12, 24 or 48 h following surgery [3H]2-deoxyglucose [( 3H]2-DG) was administered to gauge patterns of glucose uptake. In a second study, [3H]leucine was injected to assess patterns of protein synthesis. Autoradiographs were then subjected to quantitative analyses. As early as 1 h following occlusion reduced 2-DG uptake was observed in many bulb regions. By 24 h, leucine incorporation was also uniformly diminished. While 2-DG uptake remained suppressed 48 h after deprivation, levels of amino acid incorporation returned to normal patterns in most laminae, with the exception of the mitral cell layer, where increased uptake was encountered. To evaluate whether the effects were developmental by nature a group of P40-P45 animals treated similarly were also examined. While 24 h of deprivation impaired 2-DG uptake in older animals, no alterations in amino acid incorporation were observed. The results indicate that early odor deprivation has rapid and specific effects on cellular functioning within the developing olfactory bulb.

Animals↗

PubMed1989

Prenatal neurogenesis in the telencephalon of the precocial mouse Acomys cahirinus.

[3H]thymidine autoradiography was employed to examine the times of formation of the major neuronal classes in the forebrain of the precocial mouse Acomys cahirinus. Dams received 3 thymidine injections over a 24 h period on either embryonic Day 14, 18, 20, 22, 29 or 36. Age-related changes in the distribution and number of heavily labeled cells were noted. Acomys exhibited later onset and more protracted periods of cell generation than the phylogenetically related, altricial laboratory rat or mouse, indicating considerable differences in patterns of early growth between the species. Understanding the factors responsible for these differences could lend important insights into evolutionary mechanisms involved in the process of speciation.

Animals↗

PubMed1989

Early postnatal cellular proliferation and survival in the olfactory bulb and rostral migratory stream of normal and unilaterally odor-deprived rats.

Unilateral naris occlusion in rats on postnatal Day 1 results in dramatic decreases in the size of specific olfactory bulb cell populations when pups are examined 30 days later (Frazier and Brunjes: J. Comp. Neurol. 269: 355-370, '88). The observed reductions must result from alterations in cell proliferation and/or survival, alternatives examined in the present study. During early postnatal development, most cells destined for the bulb are produced in regions caudal to the structure and migrate to the bulb in the massive rostral migratory stream. The dynamics of the stream were examined in both normal rats and pups with a single naris closed on Day 1. 3H-thymidine injections were made on postnatal Days 2, 5, 10, 20, and 30. Groups of pups were killed 2 hours later to assess patterns of proliferation and 24 hours later to gauge initial stages of migration. A gradient of labeled cells was observed in the stream, with higher levels occurring at more caudal locations. The supply of cells to the bulb peaked on Day 5 and was still substantial as late as Day 30. The deprivation procedure did not affect patterns of cell labeling at any stage tested, indicating the procedure does not affect early cellular proliferation. A third group of pups was examined 30 days after thymidine injection to assess both time of cell origin and survival rates. Dark granule cells and glia in the granule cell layer were produced at a consistent rate until Day 20 with cells added during the period evenly spread throughout the layer. Light granule and periglomerular cell production decreased dramatically after P5. Thirty days after injections on P2, fewer labeled dark granule cells and their associated glia were found in deprived bulbs, indicating that enhanced cell death plays a major role in the deprivation-induced decrease in cell number.

Animals↗

PubMed1989

A comparative study of prenatal development in the olfactory bulb, neocortex and hippocampal region of the precocial mouse Acomys cahirinus and rat.

Unlike the remainder of the rodent subfamily Muridae, Acomys cahirinus (the 'spiny' mouse) is born in a precocial state after a long (39 day) gestation. In this paper, the development of the olfactory bulb, neocortex and hippocampal formation of Acomys from prenatal days 14-34 was examined and the rate of maturation compared with that of its cousin, the laboratory rat (Rattus norvegicus). At the earliest stages examined, Acomys was approximately 2 days less mature than the same post-conception aged rat. The difference between the two species increased: Acomys at 28 days postconception resembled the 22-day rat. By the end of gestation, Acomys and the rat were in a relatively similar developmental state. Therefore, Acomys exhibits a quite different timetable of early maturation which includes a protracted period of relatively slow growth during mid-gestation. As such, it offers many benefits as a subject for studies of both early ontogenesis and the mechanisms which result in species differences.

Animals↗